LOCUS       sequence-272928-        5603 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GST-tagged 4xCTD heptapeptide repeat in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5603)
  AUTHORS   Castaneda AF, Didychuk AL, Louder RK, McCollum CO, Davis ZH, Nogales
            E, Glaunsinger BA
  TITLE     The gammaherpesviral TATA-box-binding protein directly interacts 
            with the CTD of host RNA Pol II to direct late gene transcription.
  JOURNAL   PLoS Pathog. 2020 Sep 4;16(9):e1008843. doi: 
            10.1371/journal.ppat.1008843. eCollection 2020 Sep.
  PUBMED    32886723
REFERENCE   2  (bases 1 to 5603)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS 
            Pathog."; date: "2020-09-4"; volume: "16(9):e1008843. doi"; pages: "
            10.1371/journal.ppat.1008843. eCollection 2020 Sep"
COMMENT     Sequence Label: pQLink-GST-4xCTD
FEATURES             Location/Qualifiers
     source          1..5603
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..15,16..21,22..38,39..45)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    21..37
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    53..69
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             88..99
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             106..759
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     717..739
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     protein_bind    781..805
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             805..825
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(929..953)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      975..1069
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1163..1822
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1229..1248)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1887..1973
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2066..3148)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     3110..3129
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3149..3226)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(3310..3332)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3418..3558
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3473..3492
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3573..3590)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3744..4332)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3824..3843)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4503..5294,5295..5363))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5126..5145
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5364..5468)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5536..5554
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcataaaaaa tttatttgct ttgtgagcgg ataacaatta taatagattc aattgtgagc
       61 ggataacaat ttcacacaga attcattaaa gaggagaaat tacatatgtc ccctatacta
      121 ggttattgga aaattaaggg ccttgtgcaa cccactcgac ttcttttgga atatcttgaa
      181 gaaaaatatg aagagcattt gtatgagcgc gatgaaggtg ataaatggcg aaacaaaaag
      241 tttgaattgg gtttggagtt tcccaatctt ccttattata ttgatggtga tgttaaatta
      301 acacagtcta tggccatcat acgttatata gctgacaagc acaacatgtt gggtggttgt
      361 ccaaaagagc gtgcagagat ttcaatgctt gaaggagcgg ttttggatat tagatacggt
      421 gtttcgagaa ttgcatatag taaagacttt gaaactctca aagttgattt tcttagcaag
      481 ctacctgaaa tgctgaaaat gttcgaagat cgtttatgtc ataaaacata tttgaatggt
      541 gatcatgtaa cccatcctga cttcatgttg tatgacgctc ttgatgttgt tttatacatg
      601 gacccaatgt gcctggatgc gttcccaaaa ttagtttgtt ttaaaaaacg tattgaagct
      661 atcccacaaa ttgataagta cttgaaatcc agcaagtata tagcatggcc tttgcagggc
      721 tggcaagcca cgtttggtgg tggcgaccat cctccaaaat cggatctggg tgctgcaggt
      781 acaagtttgt acaaaaaagc aggcgagaat ctttattttc agggatccta tagccccacg
      841 agcccttctt actctccgac ctcaccatcc tattcgccta ctagcccgag ttacagtccc
      901 acatctccgt cctaggcggc cgcctaggac ccagctttct tgtacaaagt ggtaagctta
      961 attagctgag cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga
     1021 tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc caagtaacaa
     1081 caccatttaa atggagtggt tacaaatgga gtggttaatt aaggctagct tggcgagatt
     1141 ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat
     1201 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta
     1261 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca
     1321 caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt
     1381 tcgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac
     1441 cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt
     1501 ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta
     1561 tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt
     1621 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat
     1681 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca
     1741 tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga
     1801 tgagtggcag ggcggggcgt aattttttta aggcagttat tggtgccctt aaacgcctgg
     1861 ggtaatgact ctctagcttg aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     1921 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccct
     1981 ctagattacg tgcagtcgat gataagctgt caaacatgag aattgtgcct aatgagtgag
     2041 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2101 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     2161 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
     2221 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
     2281 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
     2341 ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
     2401 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     2461 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     2521 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
     2581 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
     2641 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
     2701 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     2761 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     2821 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     2881 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     2941 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     3001 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     3061 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     3121 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     3181 gctatcatgc cataccgcga aaggttttgc accattcgat ggtgtcggaa tttcgggcag
     3241 cgttgggtcc tggccacggg tgcgcatgat ctagagctgc ctcgcgcgtt tcggtgatga
     3301 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     3361 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     3421 agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca
     3481 gagcagattg tactgagagt gcaccacatg cggtgtgaaa taccgcacag atgcgtaagg
     3541 agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3601 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3661 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3721 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3781 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3841 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3901 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3961 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4021 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4081 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4141 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     4201 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4261 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4321 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4381 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4441 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4501 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4561 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4621 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4681 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4741 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4801 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4861 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4921 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4981 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5041 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5101 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5161 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5221 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5281 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5341 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5401 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5461 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     5521 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcacct cgagcttaat
     5581 taacaacacc atttgtcgag aaa
//
